Refinement and Applications of Light Controlled Biological Processes
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Abstract
For understanding of biological processes, a spatiotemporal control over these processes is needed. Controlling these processes using light can give us the advantage of a precise control over spacing, timing and degree of biological functions. Thus, biological processes can be turned on/off using an external light source. Multiple groups have worked on controlling cellular processes using photo-caged biomolecules. In this dissertation work, several approaches to control biological processes using caged molecules are described. In addition, applications and refinement of these processes are also discussed in detail. First chapter of this dissertation deals with various strategies used to enhance light activated RNA interference (LARI). LARI is a technique developed in our laboratory to control spacing, timing and degree of gene expression using light. We have demonstrated that incorporation of just two phosphorothioate linkages in strands of double stranded RNA leads to a significant increase in the window of toggled expression in cells. In second chapter, we have demonstrated that cells can be successfully patterned by spatially manipulating biology as oppose to spatial manipulation of the surfaces on which cells are grown. To pursue this, we incorporated PTEN siRNA into the LARI process. Additionally, photo-caged apoptosis inducers were also evaluated for the purpose of patterning the cells. In final chapters of this dissertation, we discussed about construction of light activated siRNA nanoparticles and synthesis of light activated proteins. We have successfully demonstrated caging of proteins using commercially available photo-labile group. Furthermore, we have shown that with the use of these caged proteins, the activity of protein can be spatiotemporally controlled using light.
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Introduction and literature review -- Approaches to improve LARI -- Patterning of cells by patterning biology -- Light activated siRNA -- Light activated proteins -- Appendix
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Ph.D.